以雪莲叶片为外植体,探讨了卡那霉素(kanamycin,Kan)、潮霉素(hygromycin B,Hyg)、羧苄青霉素(car-benidillin,Car)3种抗生素对雪莲愈伤组织诱导、生长及分化的影响,以确定农杆菌介导的遗传转化研究中筛选剂和抑菌剂的最适浓度。结果表明:40mg/L的卡那霉素已抑制雪莲愈伤组织生长,当卡那霉素为50mg/L时,愈伤组织的生长基本停止;8.0mg/L潮霉素能够有效抑制雪莲愈伤组织的生长,当潮霉素为20mg/L时则生长的愈伤组织块较小、褐化、甚至死亡。同时,低浓度(0.5~2.0mg/L)的潮霉素可以提高雪莲愈伤组织的分化率;作为农杆菌抑菌剂,不同浓度羧苄青霉素对雪莲愈伤组织生长的影响差异极显著,当羧苄青霉素的浓度超过400mg/L时对雪莲愈伤组织的出愈及生长均有明显的抑制作用。
The effects of different antibiotics(kanamycin,hygromycin B and carbenidillin) on induction and differentiation of callus(Saussurea involucrata),was discussed in this study and S.involucrata leaves were used as explants.A doses strategy of selection and bacterium inhibition during agrobacter mediated genetic transformation was established.The result showed that the kanamycin of 40 mg/L was significantly influenced on callus growth,when it was 50 mg/L,the growth of callus was inhibited completely;the hygromycin of 8 mg/L was the best concentration for inhibiting the callus growth.The callus were geatly damaged and it's characters were small,browning and dying with the concentration of hygromycin up to 20 mg/L.Meanwhile,the low concentration(0.5~2.0 mg/L) of hygromycin can enhance the rate of callus differentiation.Carbenidillin is an antibacterial agent of Agrobacterium and carbenidillin had apparently influnced on growth of callus at different concentrations.When the concentration of carbenidillin was greater than 400 mg/L,the growth of callus was effected observably.This paper may provided a theoretical for S.involucrata genetic transformation system.